Problems of Forensic Sciences 2010 Vol. 82 (LXXXII) 161-172

APPLICATION OF THE EMIT AND HPLC-DAD METHODS TO DETERMINATION OF THE CONCENTRATION OF BENZODIAZEPINES IN THE SERUM OF POISONED PATIENTS

Ewa GOMÓŁKA1, 3, Jolanta WILIMOWSKA1, Adrianna SULKA1, Tomasz GAWLIKOWSKI2, Agnieszka MORAWSKA1
1
Department of Analytical Toxicology and Therapeutic Drug Monitoring, Collegium Medicum, Jagiellonian University, Kraków
2Toxicology Clinic, Faculty of Toxicology and Environmental Diseases, Collegium Medicum, Jagiellonian University, Kraków
3Toxicological Laboratory, Department of Laboratory Diagnostics, L. Rydygier’s Hospital, Kraków

Streszczenie
About 300 cases of benzodiazepine intoxications per year are diagnosed in the Toxicology Clinic, Collegium Medicum, Jagiellonian University in Kraków. Therefore, there is a need for rapid and reliable methods that allow determination of the concentrations of these drugs in body fluids. The aim of this study was to estimate the usefulness of the enzyme multiplied immunoassay technique (EMIT) for determination of benzodiazepines in serum. Blood was collected from patients poisoned with benzodiazepines diagnosed in the Laboratory of Analytical Toxicology and Drug Monitoring UJ CM. EMIT (Viva-E, Siemens) and high pressure liquid chromatography with diode array detection (HPLC-DAD) (LaChrom, Merck-Hitachi) were used. The results of benzodiazepines concentrations in serum obtained by the EMIT method in the group of patients ranged from a few to over 5000 ng/ml. The presence of benzodiazepines (parent drugs and their metabolites) was confirmed in all samples by the HPLC-DAD method. Diazepam, estazolam, and nordiazepam (accounting together for more than 60% of the ¬results) and oxazepam, alprazolam, clonazepam, lorazepam, and temazepam were detected most often. The specificity of the analytical method regarding the measured substance and its therapeutic and toxic serum ranges must be considered in interpretation of ¬results obtained by the EMIT method. Confirmation analysis by a reference method should be considered in questionable cases.

Słowa kluczowe
Benzodiazepines; Poisoning; EMIT; HPLC-DAD.

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